Differential Cleaving of Specific Substrates for Cathepsin-Like Activity Shows Cysteine and Serine Protease Activities and a Differential Profile Between<i>Anisakis simplex s.s.</i>and<i>Anisakis pegreffii</i>, Sibling Species Major Etiologic Agents of Anisakiasis

Torralbo-Ramírez, Verónica; Molina-Fernández, Dolores; Malagón, David; Benı́tez, Rocı́o; Adroher, Francisco Javier

doi:10.1089/fpd.2019.2633

Cited by 4 publications

(4 citation statements)

References 49 publications

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“…Additional, rTsSPc hydrolyzed collagen I was dose-dependent at pH 8.0. The results were consistent with the optimal pH values of most serine proteases from other parasites previously reported [ 70 ]. Serine proteases in Fasciola hepatica miracidia hydrolyzed azocasein with optimum activity at pH 8.0.…”

Section: Discussionsupporting

confidence: 92%

“…To assay the enzymatic activity of rTsSPc, the serially diluted rTsSPc (0, 0.01, 0.02, 0.04, 006, 0.08, 0.10 μg/μl) was first pre-incubated at 37 ˚C for 15 min in various pH buffer (pH 3.0-5.0 sodium acetate, pH 6.0-7.0 sodium phosphate, pH 8.0-9.0 Tris-HCl, and pH 10.0-10.5 sodium bicarbonate). Subsequently, the substrate N-benzoyl-L-arginine-ethylester (BAEE; BBI, Shanghai, China) was added to the reaction mixture and incubated at different temperatures (20,30,37,40,50,60,70, and 80 ˚C) for 15 min, and the absorbance at 253 nm was measured using a spectrophotometer [48]. To evaluate the effect of different metal ions on rTsSPc enzyme activity, metal ions (Ca 2+ , Fe 2+ , Zn 2+ , Mg 2+ , Ni 2+ , and Mn 2+ ) were added to the reaction system [12,49].…”

Section: Assay Of Rtsspc Enzyme Activity By Using Substrate Baeementioning

confidence: 99%

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Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium

Song,

Zhang,

Wang

et al. 2023

PLoS Negl Trop Dis

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Background A novel serine proteinase of Trichinells spiralis (TsSPc) has been identified in the excretion/secretion (ES) antigens, but its role in larval invasion is unclear. The aim of this study was to clone and express TsSPc, identify its biological and biochemical characteristics, and investigate its role on larval invasion of gut epithelium during T. spiralis infection. Methodology/Principal findings TsSPc has a functional domain of serine proteinase, and its tertiary structure consists of three amino acid residues (His88, Asp139 and Ser229) forming a pocket like functional domain. Recombinant TsSPc (rTsSPc) was expressed and purified. The rTsSPc has good immunogenicity. On Western blot analysis, rTsSPc was recognized by infection serum and anti-rTsSPc serum, natural TsSPc in crude and ES antigens was identified by anti-rTsSPc serum. The results of qPCR, Western blot and indirect immunofluorescence test (IIFT) showed that TsSPc was expressed at diverse stage worms, and mainly localized at cuticle, stichosome and intrauterine embryos of this nematode. The rTsSPc had enzymatic activity of native serine protease, which hydrolyzed the substrate BAEE, casein and collagen I. After site directed mutation of enzymatic active sites of TsSPc, its antigenicity did not change but the enzyme activity was fully lost. rTsSPc specifically bound to intestinal epithelium cells (IECs) and the binding sites were mainly localized in cell membrane and cytoplasm. rTsSPc accelerated larval invasion of IECs, whereas anti-rTsSPc antibodies and TsSPc-specific dsRNA obviously hindered larval invasion. Conclusions TsSPc was a surface and secretory proteinase of the parasite, participated in larval invasion of gut epithelium, and may be considered as a candidate vaccine target molecule against Trichinella intrusion and infection.

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Section: Discussionsupporting

confidence: 92%

Section: Assay Of Rtsspc Enzyme Activity By Using Substrate Baeementioning

confidence: 99%

Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium

Song,

Zhang,

Wang

et al. 2023

PLoS Negl Trop Dis

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“…Additionally, endopeptidases, like the serine proteases detected in this study, play a significant role in the lifecycle of the parasite and in the pathogen‒host relationship that could be related to virulence, invasion of host tissues, and/or intracellular digestion ( File S2 ) [ 49 , 50 ]. Moreover, the previously mentioned putative resistance of Fasciola hepatica to triclabendazole, characterized by sublethal activity, was indicated by the presence of cathepsin L, a peptidase [ 46 ].…”

Section: Resultsmentioning

confidence: 99%

Comparative Proteomics Analysis of Anisakis simplex s.s.—Evaluation of the Response of Invasive Larvae to Ivermectin

Polak

Łopieńska–Biernat

Stryiński

et al. 2020

Genes

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Ivermectin (IVM), an antiparasitic drug, has a positive effect against Anisakis simplex s.s. infection and has been used for the treatment and prevention of anisakiasis in humans. However, the molecular mechanism of action of IVM on A. simplex s.s. remains unknown. Herein, tandem mass tag (TMT) labeling and extensive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis were used to identify the effect of IVM on the proteome of A. simplex s.s. in vitro. During the study, 3433 proteins, of which 1247 had at least two protein unique peptides, were identified. Comparative proteomics analysis revealed that 59 proteins were differentially regulated (DRPs) in IVM-treated larvae, of which 14 proteins were upregulated and 38 were downregulated after 12 h of culture, but after 24 h, 12 proteins were upregulated and 22 were downregulated. The transcription level of five randomly selected DRPs was determined by real-time PCR as a supplement to the proteomic data. The functional enrichment analysis showed that most of the DRPs were involved in oxidoreductase activity, immunogenicity, protein degradation, and other biological processes. This study has, for the first time, provided comprehensive proteomics data on A. simplex s.s. response to IVM and might deliver new insight into the molecular mechanism by which IVM acts on invasive larvae of A. simplex s.s.

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“…simplex s.s. seems more pathogenic than A. pegreffii (Jeon and Kim, 2015;Quiazon et al, 2011;Romero et al, 2013;Suzuki et al, 2010). However, this requires confirmation, while possible differences between the two should be identified (Cavallero et al, 2020(Cavallero et al, , 2018Llorens et al, 2018;Molina-Fernández et al, 2019;Torralbo-Ramírez et al, 2019), including allergens (Arcos et al, 2014;Baird et al, 2016), as they may affect diagnostic methods. As it has also been suggested that anisakiasis constitutes a risk factor for stomach and colon cancers (Corcuera et al, 2018;García-Pérez et al, 2015), studies to determine its relationship with these and other diseases are especially relevant.…”

Section: Future Studies and Research Neededmentioning

confidence: 99%

Anisakiasis and Anisakis: An underdiagnosed emerging disease and its main etiological agents

Adroher

Benítez-Rodríguez

2020

Research in Veterinary Science

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Differential Cleaving of Specific Substrates for Cathepsin-Like Activity Shows Cysteine and Serine Protease Activities and a Differential Profile BetweenAnisakis simplex s.s.andAnisakis pegreffii, Sibling Species Major Etiologic Agents of Anisakiasis

Cited by 4 publications

References 49 publications

Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium

Molecular characterization of a novel serine proteinase from Trichinella spiralis and its participation in larval invasion of gut epithelium

Comparative Proteomics Analysis of Anisakis simplex s.s.—Evaluation of the Response of Invasive Larvae to Ivermectin

Anisakiasis and Anisakis: An underdiagnosed emerging disease and its main etiological agents

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